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Rapid diagnosis of soybean anthracnose caused by Colletotrichum truncatum using a loop-mediated isothermal amplification (LAMP) assay

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Abstract

A loop-mediated isothermal amplification (LAMP) assay that directly detects Colletotrichum truncatum in diseased soybean tissues is described, thus allowing rapid diagnosis of soybean anthracnose. Using the target gene Rpb1 (that codes for the large subunit of RNA polymerase II), we designed and screened a set of species-specific primers allowing amplification at 62 °C over 70 min. After addition of SYBR Green I to the LAMP reaction products, a yellow-green color (visible to the unaided eye) developed only in the presence of C. truncatum. The detection limit of the LAMP assay was 100 pg (per μL genomic DNA). The Rpb1-Ct-LAMP assay has been successfully used to diagnose soybean anthracnose in field samples collected from Jiangsu, Anhui and Hubei provinces of China, and to detect C. truncatum in soybean seeds from farmers’ markets. Our results show that the Rpb1-Ct-LAMP assay is a useful and convenient method for detecting C. truncatum, and thus for diagnosis of soybean anthracnose.

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Acknowledgements

This research was supported by the National High-Tech R&D Program (program 863) (grant no. 2012AA101501), the National Department Public Benefit Research Foundation (grant no. 200903004), the Chinese National Science Foundation Committee (project 31225022), and public sector research funding (grant no. 201303018)

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Authors and Affiliations

  1. Department of Plant Pathology, College of Plant Protection and Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing Agricultural University, Nanjing, 210095, China

    Qing Tian, Chenchen Lu, Shuaishuai Wang, Qin Xiong, Haifeng Zhang, Yuanchao Wang & Xiaobo Zheng

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  1. Qing Tian
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  2. Chenchen Lu
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  3. Shuaishuai Wang
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  4. Qin Xiong
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  5. Haifeng Zhang
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  6. Yuanchao Wang
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  7. Xiaobo Zheng
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Correspondence to Xiaobo Zheng.

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Tian, Q., Lu, C., Wang, S. et al. Rapid diagnosis of soybean anthracnose caused by Colletotrichum truncatum using a loop-mediated isothermal amplification (LAMP) assay. Eur J Plant Pathol 148, 785–793 (2017). https://doi.org/10.1007/s10658-016-1132-2

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